The pharmacology and molecular pharmacology of various gastrointestinal (GI) peptides are being investigated. Two peptide receptor families investigated during the year are those for bombesin- (Bn) related peptides and for VIP-related peptides. Bn-related peptides (GRP, NMB) interact primarily with two distinct receptors (GRP-R, NMB-R) to mediate a number of effects in the GI tract and central nervous sytem (CNS). The aim of one aspect of this project is to understand the pharmacology, molecular pharmacology, and cell biology of these receptors. To identify key amino acids responsible for selective high affinity binding of various agonists for different members of the bombesin receptor family we used an analysis of the structural homologies of different receptors of this family and site-directed mutagenesis. Between the GRP receptor and BRS-3 recptor which different by >1000 in their affinity for bombesin (BN) we found 14 important amino acid differences which were then substituted in each receptor. Molecular modeling and pharmacolgy studies showed 7 of these were important for high affinity for Bn and contibuted to the selectivity of these receptors demonstrating this is a useful approach to identify amino acid important for selectivity and affinity. A second aspect of our studies is to develop selective receptor ligands for Bn or VIP receptor subtypes that could be metabolically stable. To accomplish this for the human BRS-3 receptor which has no selective ligands, we made substitutions of various amino acids in a nonselective ligand that we had discovered that interacted with with this receptor with high affinity. One analog dTyr6, Apa-4 Cl, Phe13, Nle14-Bn (6-14) was found to have a marked enhanced selectivity for the human BRS3 receptor over the other receptors of this family and should be useful to investigate its role in physiological and pathological processes. Lastly using our previous ligand structure function studies we were able to construct simplified analogs of the 28 amino acide peptide VIP that were metabolically stable and had high affinity and selectivity for VPAC1 receptors.